Figure 4: Nestin⁺ cells in the aortic adventitia and media express Mcp1 in mice and humans.

Immunofluorescence of sections from (a,b,f) thoracic aorta and (c) aortic valves from Nes-Gfp mice using antibodies against (a,b) GFP and (c) CD31 (red). Note that nestin⁺ cells are mainly distributed in the adventitia (b) or in close proximity of valve endothelial cells (c). Inset, high magnification of the aortic leaflets. (d,e) Immunofluorescence of representative control section from WT thoracic aorta using antibodies against GFP (red). The absence of red signal demonstrates the specificity of the staining. (b,e) The autofluorescence of the smooth muscle layer is shown in green. (f) Negative control immunofluorescence on Nes-gfp aorta using control IgG (top panels) and/or secondary antibodies (bottom panels). (b,c,e,f) Nuclei were counterstained with DAPI (blue). (g–j) Immunofluorescence of (g–i) NESTIN and (h–j) MCP1 in consecutive cross sections from human (g,h) coronary and (i,j) carotid artery samples. Red arrowheads depict cells that express both proteins in consecutive sections. (a,b,d–f,g–j) Scale bars, 50 μm, (c) 100 μm.